An image analysis method includes acquiring images of spatially different analysis regions. Each of the images of the analysis regions is constituted by pixels including a plurality of data acquired simultaneously or time-serially. The method further includes obtaining a cross-correlation between two analysis regions by using data of pixels of images of the analysis regions.
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1. An image analysis method comprising: acquiring images of spatially different analysis regions, each of the images of the analysis regions being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; and obtaining a cross-correlation between two analysis regions by using data of pixels of the images of the analysis regions, wherein each image of the analysis regions is a two-dimensional image, the two analysis regions, between which the cross-correlation is obtained, comprise an analysis region 1 and an analysis region 2 , and the obtaining the cross-correlation performs correlation calculation by using equation (1) and fitting for the correlation calculation result by using equation (2) to obtain the cross-correlation between the two-dimensional analysis regions: G s ( ξ , ψ ) = ∑ I 1 ( x , y ) * I 2 ( x + ξ , y + ψ ) / M 12 ( ∑ I 1 ( x , y ) / M 1 ) ( ∑ I 2 ( x , y ) / M 2 ) ( 1 ) where G s is a spatial cross-correlation value of RICS, I 1 is data of a pixel of an image of the analysis region 1 , I 2 is data of a pixel of an image of the analysis region 2 , x and y are the spatial coordinates of a measurement point, ξ and ψ are the changes of the spatial coordinates from the measurement point, M 12 is the number of times of sum-of-product calculation of data of the analysis region 1 and the analysis region 2 , M 1 is the total number of data of the analysis region 1 , and M 2 is the total number of data of the analysis region 2 G s ( ξ , ψ ) = S ( ξ , ψ ) * G ( ξ , ψ ) S ( ξ , ψ ) = exp ( - 1 2 * [ ( 2 ξ δ r W 0 ) 2 + ( 2 ψ δ r W 0 ) 2 ] ( 1 + 4 D ( τ p ξ + τ l ψ ) W 0 2 ) ) G ( ξ , ψ ) = 1 N ( ( 1 + 4 D ( τ p ξ + τ l ψ ) W 0 2 ) - 1 * ( 1 + 4 D ( τ p ξ + τ l ψ ) W Z 2 ) - 1 / 2 ) ( 2 ) where Gs is a spatial correlation value of RICS, S is an influence of a scan in RICS analysis, G is an influence of a time delay in RICS analysis, D is a diffusion constant, δ r is a pixel size, N is the number of molecules, ξ and ψ are the changes of spatial coordinates from a measurement point, W 0 is a radius of an excitation laser beam in a lateral direction, W z is a radius of an excitation laser beam in a longitudinal direction, τ p is a pixel time, and τ l is a line time.
2. The method of claim 1 , wherein the acquiring images of analysis regions comprising: acquiring an image of an observation region including the analysis regions, the image of the observation region being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; setting the analysis regions for the image of the observation region; and extracting data of pixels corresponding to the analysis regions from the image of the observation region.
3. The method of claim 2 , wherein the acquiring the image of the observation region acquires images of frames of the observation region.
4. The method of claim 3 , wherein the obtaining the cross-correlation obtains the cross-correlation by using any one of a fluorescence intensity, a pixel time, a line time, a frame time, a pixel positional relationship, a pixel size, and a statistical value thereof.
5. The method of claim 1 , wherein the acquiring the images of the analysis regions acquires images of the analysis regions respectively by using optical systems.
6. The method of claim 5 , wherein the acquiring the images of the analysis regions simultaneously acquires images of the analysis regions.
7. The method of claim 5 , wherein the acquiring the images of the analysis regions acquires images of frames of the analysis regions.
8. The method of claim 7 , wherein the obtaining the cross-correlation obtains the cross-correlation by using any one of a fluorescence intensity, a pixel time, a line time, a frame time, a pixel positional relationship, a pixel size, and a statistical value thereof.
9. The method of claim 1 , wherein the obtaining the cross-correlation obtains the cross-correlation by using any one of an average value, a maximum value, a minimum value, a relative difference, and an absolute difference of the data.
10. The method of claim 1 , wherein the obtaining the cross-correlation obtains the cross-correlation by using reconstructed data obtained by reconstructing the data.
11. An image analysis method comprising: acquiring images of spatially different analysis regions, each of the images of the analysis regions being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; and obtaining a cross-correlation between two analysis regions by using data of pixels of the images of the analysis regions, wherein each image of the analysis regions is a three-dimensional image, the two analysis regions, between which the cross-correlation is obtained, are an analysis region 1 and an analysis region 2 , and the obtaining the cross-correlation performs correlation calculation by using equation (3) and fitting for the correlation calculation result by using equation (4) to obtain the cross-correlation between the three-dimensional analysis regions: G s ( ξ , ψ , η ) = ∑ I 1 ( x , y , z ) * I 2 ( x + ξ , y + ψ , z + η ) / M 12 ( ∑ I 1 ( x , y , z ) / M 1 ) ( ∑ I 2 ( x , y , z ) / M 2 ) ( 3 ) where Gs is a spatial cross-correlation value of RICS, I 1 is data of a pixel of an image of the analysis region 1 , I 2 is data of a pixel of an image of the analysis region 2 , x, y, and z are the spatial coordinates of a measurement point, ξ, ψ, and η are the changes of the spatial coordinates from the measurement point, M 12 is the number of times of sum-of-product calculation of data of the analysis region 1 and the analysis region 2 , M 1 is the total number of data of the analysis region 1 , and M 2 is the total number of data of the analysis region 2 G s ( ξ , ψ , η ) = S ( ξ , ψ , η ) * G ( ξ , ψ , η ) S ( ξ , ψ , η ) = exp ( - 1 2 * [ ( 2 ξ δ r W 0 ) 2 + ( 2 ψ δ r W 0 ) 2 + ( 2 η δ r W 0 ) 2 ] ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W 0 2 ) ) G ( ξ , ψ , η ) = 1 N ( ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W 0 2 ) - 1 * ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W Z 2 ) - 1 / 2 ) ( 4 ) where Gs is a spatial correlation value of RICS, S is an influence of a scan in RICS analysis, G is an influence of a time delay in RICS analysis, D is a diffusion constant, δ r is a pixel size, N is the number of molecules, ξ, ψ, and η are the changes of spatial coordinates, W 0 is a radius of an excitation laser beam in a lateral direction, W z is a radius of an excitation laser beam in a longitudinal direction, τ p is a pixel time, τ l is a line time, and τ f is a frame time.
12. An image analysis apparatus comprising: an analysis region image acquisition unit configured to acquire images of spatially different analysis regions, each of the images of the analysis regions being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; and a correlation analysis unit configured to obtain a cross-correlation between two analysis regions by using data of pixels of the images of the analysis regions, wherein each image of the analysis regions is a two-dimensional image, the two analysis regions, between which the cross-correlation is obtained, are an analysis region 1 and an analysis region 2 , and the correlation analysis unit performs correlation calculation by using equation (5) and fitting for the correlation calculation result by using equation (6) to obtain the cross-correlation between the two-dimensional analysis regions: G s ( ξ , ψ ) = ∑ I 1 ( x , y ) * I 2 ( x + ξ , y + ψ ) / M 12 ( ∑ I 1 ( x , y ) / M 1 ) ( ∑ I 2 ( x , y ) / M 2 ) ( 5 ) where G s is a spatial cross-correlation value of RICS, I 1 is data of a pixel of an image of the analysis region 1 , I 2 is data of a pixel of an image of the analysis region 2 , x and y are the spatial coordinates of a measurement point, ξ and ψ are the changes of the spatial coordinates from the measurement point, M 12 is the number of times of sum-of-product calculation of data of the analysis region 1 and the analysis region 2 , M 1 is the total number of data of the analysis region 1 , and M 2 is the total number of data of the analysis region 2 G s ( ξ , ψ ) = S ( ξ , ψ ) * G ( ξ , ψ ) S ( ξ , ψ ) = exp ( - 1 2 * [ ( 2 ξ δ r W 0 ) 2 + ( 2 ψ δ r W 0 ) 2 ] ( 1 + 4 D ( τ p ξ + τ l ψ ) W 0 2 ) ) G ( ξ , ψ ) = 1 N ( ( 1 + 4 D ( τ p ξ + τ l ψ ) W 0 2 ) - 1 * ( 1 + 4 D ( τ p ξ + τ l ψ ) W Z 2 ) - 1 / 2 ) ( 6 ) where G s is a spatial correlation value of RICS, S is an influence of a scan in RICS analysis, G is an influence of a time delay in RICS analysis, D is a diffusion constant, δ r is a pixel size, N is the number of molecules, ξ and ψ are the changes of spatial coordinates from a measurement point, W 0 is a radius of an excitation laser beam in a lateral direction, W z is a radius of an excitation laser beam in a longitudinal direction, τ p is a pixel time, and τ l is a line time.
13. The apparatus of claim 12 , wherein the analysis region image acquisition unit comprising: an observation region image acquisition unit configured to acquire an image of an observation region including the analysis regions, the image of the observation region being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; an analysis region setting unit configured to set the analysis regions for the image of the observation region; and a data extraction unit configured to extract data of pixels corresponding to the analysis regions from the image of the observation region.
14. The apparatus of claim 13 , wherein the observation region image acquisition unit acquires images of frames of the observation region.
15. The apparatus of claim 14 , wherein the correlation analysis unit obtains the cross-correlation by using any one of a fluorescence intensity, a pixel time, a line time, a frame time, a pixel positional relationship, a pixel size, and a statistical value thereof.
16. The apparatus of claim 12 , wherein the analysis region image acquisition unit includes optical systems configured to respectively acquire images of the analysis regions.
17. The apparatus of claim 16 , wherein the analysis region image acquisition unit simultaneously acquires images of the analysis regions.
18. The apparatus of claim 16 , wherein the analysis region image acquisition unit acquires images of frames of the analysis regions.
19. The apparatus of claim 16 , wherein the correlation analysis unit obtains the cross-correlation by using any one of a fluorescence intensity, a pixel time, a line time, a frame time, a pixel positional relationship, a pixel size, and a statistical value thereof.
20. The apparatus of claim 12 , wherein the correlation analysis unit obtains the cross-correlation by using any one of an average value, a maximum value, a minimum value, a relative difference, and an absolute difference of the data.
21. The apparatus of claim 12 , wherein the correlation analysis unit obtains the cross-correlation by using reconstructed data obtained by reconstructing the data.
22. An image analysis apparatus comprising: an analysis region image acquisition unit configured to acquire images of spatially different analysis regions, each of the images of the analysis regions being constituted by pixels including a plurality of data acquired simultaneously or time-serially, respectively; and a correlation analysis unit configured to obtain a cross-correlation between two analysis regions by using data of pixels of the images of the analysis regions, wherein each image of the analysis region is a three-dimensional image, the two analysis regions, between which the cross-correlation is obtained, are an analysis region 1 and an analysis region 2 , and the correlation analysis unit performs correlation calculation by using equation (7) and fitting for the correlation calculation result by using equation (8) to obtain the cross-correlation of the three-dimensional analysis regions: G s ( ξ , ψ , η ) = ∑ I 1 ( x , y , z ) * I 2 ( x + ξ , y + ψ , z + η ) / M 12 ( ∑ I 1 ( x , y , z ) / M 1 ) ( ∑ I 2 ( x , y , z ) / M 2 ) ( 7 ) where G s is a spatial cross-correlation value of RICS, I 1 is data of a pixel of an image of the analysis region 1 , I 2 is data of a pixel of an image of the analysis region 2 , x, y, and z are the spatial coordinates of a measurement point, ξ, ψ, and η are the changes of the spatial coordinates from the measurement point, M 12 is the number of times of sum-of-product calculation of data of the analysis region 1 and the analysis region 2 , M 1 is the total number of data of the analysis region 1 , and M 2 is the total number of data of the analysis region 2 G s ( ξ , ψ , η ) = S ( ξ , ψ , η ) * G ( ξ , ψ , η ) S ( ξ , ψ , η ) = exp ( - 1 2 * [ ( 2 ξ δ r W 0 ) 2 + ( 2 ψ δ r W 0 ) 2 + ( 2 η δ r W 0 ) 2 ] ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W 0 2 ) ) G ( ξ , ψ , η ) = 1 N ( ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W 0 2 ) - 1 * ( 1 + 4 D ( τ p ξ + τ l ψ + τ f η ) W Z 2 ) - 1 / 2 ) ( 8 ) where G s is a spatial correlation value of RIGS, S is an influence of a scan in RIGS analysis, G is an influence of a time delay in RIGS analysis, D is a diffusion constant, δ r is a pixel size, N is the number of molecules, ξ, ψ, and η are the changes of spatial coordinates, W 0 is a radius of an excitation laser beam in a lateral direction, W z is a radius of an excitation laser beam in a longitudinal direction, τ p is a pixel time, τ l is a line time, and τ f is a frame time.
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June 24, 2011
December 9, 2014
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